CN111956676A - A microecological preparation containing Ginseng radix extract and probiotic bacteria, and its preparation method and application - Google Patents

A microecological preparation containing Ginseng radix extract and probiotic bacteria, and its preparation method and application Download PDF

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CN111956676A
CN111956676A CN202010859938.4A CN202010859938A CN111956676A CN 111956676 A CN111956676 A CN 111956676A CN 202010859938 A CN202010859938 A CN 202010859938A CN 111956676 A CN111956676 A CN 111956676A
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probiotic
lactobacillus
ginseng
bifidobacterium
cfu
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王苹
黄鹤
白亮
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Tianjin Modern Innovation Traditional Chinese Medicine Technology Co ltd
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Tianjin Modern Innovation Traditional Chinese Medicine Technology Co ltd
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Abstract

The invention provides a microecological preparation comprising a ginseng extract and probiotics, wherein the ginseng extract is an aqueous extract of ginseng, and the probiotics are selected from one or a combination of two or more of bifidobacteria, lactobacilli, streptococci, bacilli, clostridium butyricum and yeasts. The components of the traditional Chinese medicine ginseng can provide substrates required by growth and synthesis of active effective molecules for probiotics, and in turn, a complex enzyme system in the probiotics further improves the utilization of the traditional Chinese medicine components of the ginseng, and the traditional Chinese medicine components and the complex enzyme system can play a combined role when being used in combination.

Description

A microecological preparation containing Ginseng radix extract and probiotic bacteria, and its preparation method and application
Technical Field
The invention belongs to the technical field of traditional Chinese medicine biology, and particularly relates to a microecological preparation containing a ginseng extract and probiotics, and a preparation method and application thereof.
Background
The incidence of obesity is rising worldwide and has become a global disease that threatens human health and reduces human quality of life. Obesity, a complex disease in which genetic factors, environmental factors and lifestyle interact, increases the risk of metabolic diseases such as hyperlipidemia, type 2 diabetes, fatty liver, and the like. The fundamental cause of obesity is imbalance between intake and consumption of energy, but the cause of obesity is not completely understood, and obesity is currently considered to be a chronic disease mainly caused by the combined action of host factors and environmental factors. It is often difficult to combat obesity or metabolic diseases through reasonable dietary control and physical exercise, and the use of medication for control has side effects and causes damage to the body through long-term use.
The intestinal flora is closely related to the health of human body, and the metabolic activity of the intestinal flora influences the nutrition, metabolism, immune function and the like of a host, and is an important environmental factor influencing obesity and related metabolic diseases. Short Chain Fatty Acids (SCFA) produced by the metabolism of the gut flora have an effect on maintaining human health, metabolism and immune homeostasis. Research in recent years shows that the traditional Chinese medicine ginseng has the effects of regulating blood sugar and blood fat and promoting insulin secretion, has a good regulating effect on glycolipid metabolism, and can obviously improve obesity or metabolic syndrome. Meanwhile, researches show that the ginseng can target intestinal flora, increase energy metabolism and reduce fat accumulation by regulating the composition and the function of metabolic flora. However, the simple Chinese medicinal preparation has no obvious effect on regulating the metabolism of a human body, has an unsatisfactory effect on reducing fat accumulation, and has a limited effect on improving key beneficial molecules in the intestinal tract such as short-chain fatty acids. In addition, excessive administration of ginseng preparation increases the burden on the human body, has negative effects such as blood pressure increase, and increases the burden on the heart.
The intestinal flora is primarily bacteria, which can be roughly divided into three major groups: beneficial bacteria, harmful bacteria and neutral bacteria. Beneficial bacteria, also called probiotics, mainly comprise various bifidobacteria, lactobacilli and the like, are indispensable elements for human health, can synthesize various vitamins, participate in the digestion of food, promote intestinal tract peristalsis, inhibit the growth of pathogenic flora, decompose harmful and toxic substances and the like. Common probiotics comprise lactic acid producing strains such as bifidobacteria, lactobacilli, streptococcus and the like, and are widely applied to the aspect of digestive tract diseases, particularly intestinal tract dysfunction including constipation and diarrhea. However, probiotics have been used less frequently in obesity or metabolic diseases, because it has been found that these probiotics have a very limited ameliorating effect on obesity and metabolic diseases. The bacillus subtilis thallus can cause intestinal hypoxia in the intestinal tract and promote the growth of beneficial anaerobes as one kind of probiotics, and active substances such as subtilin, polymyxin, nystatin, gramicidin and the like generated in the growth process have obvious inhibiting effect on pathogenic bacteria or conditional pathogenic bacteria of endogenous infection. However, the use of Bacillus subtilis in obesity or metabolic diseases is still very limited.
Therefore, in the prevention and treatment of obesity and metabolic diseases, it is clearly of great significance if a microecological preparation which is highly safe, has no toxic or side effects, and can be used for a long time can be found to treat or prevent obesity and metabolic diseases by regulating the composition and function of intestinal flora.
Disclosure of Invention
To overcome the problems of the prior art, in one aspect, the present invention provides a microecological formulation comprising a ginseng extract and probiotics; wherein the Ginseng radix extract is water extract of Ginseng radix; the probiotic bacteria are selected from one or more of bifidobacteria, lactobacilli, streptococci, bacilli, clostridium butyricum and yeasts.
According to an embodiment of the invention, the probiotic bacteria are selected from one, a combination of two or more of lactobacillus acidophilus, lactobacillus rhamnosus, lactobacillus paracasei, lactobacillus casei, lactobacillus johnsonii, lactobacillus plantarum, lactobacillus fermentum, lactobacillus lactis, lactobacillus delbrueckii, lactobacillus helveticus, lactobacillus bulgaricus, lactococcus lactis diacetyl subspecies, lactococcus lactis cremoris, streptococcus thermophilus, bifidobacterium lactis, bifidobacterium animalis, bifidobacterium longum, bifidobacterium breve, bifidobacterium infantis, bifidobacterium adolescentis, lactobacillus reuteri, lactobacillus grignard, bifidobacterium bifidum, bacillus coagulans, bacillus subtilis, bacillus licheniformis, clostridium butyricum and yeasts. Preferably, one or a combination of two or more of bifidobacterium lactis, bifidobacterium animalis, bifidobacterium longum, bifidobacterium breve, bifidobacterium infantis, bifidobacterium bifidum, bacillus coagulans, bacillus subtilis and bacillus licheniformis. Most preferably, the probiotic is bacillus subtilis.
According to an embodiment of the present invention, the ginseng extract may be prepared by: extracting Ginseng radix with water under reflux, and concentrating the extractive solution to obtain Ginseng radix extract. The density of the Ginseng radix extract at 20 deg.C can be 1.0-1.4g/L, such as 1.01-1.35g/L, 1.02-1.3 g/L.
According to an embodiment of the present invention, wherein the total content of the ginsenosides Rg1 and Re in the ginseng extract is 0.5% or more, the content of the ginsenosides Rb1 is 0.2% or more; preferably, the total content of the ginsenoside Rg1 and Re in the ginseng extract is more than 0.6%, and the content of the ginsenoside Rb1 is more than 0.25%; illustratively, the total content of ginsenoside Rg1 and Re in the ginseng extract is more than 0.75%, and the content of ginsenoside Rb1 is more than 0.31%.
According to an embodiment of the invention, the microecological formulation further comprises a protectant. The protective agent is selected from one or combination of two or more of gelatin, trehalose, mannitol, vitamin C, sodium glutamate, casein, skim milk and sorbitol. Preferably, the protective agent is a mixture of gelatin, trehalose, mannitol and vitamin C in a weight ratio of gelatin to trehalose to mannitol to vitamin C of (5-50) to (5-100) to (5-50) to (0.1-2), for example, (8-30) to (8-60) to (8-30) to (0.5-1.5), illustratively 10:20:10: 1.
According to embodiments of the invention, the weight ratio of the ginseng extract to the probiotic may be (0.1-5):1, e.g. (0.2-2.5):1, (0.3-2):1, (0.4-1.8):1, (0.5-1.5):1, (0.6-1.2):1, etc.
According to an embodiment of the invention, the probiotic bacteria in the probiotic preparation have a viable count of about 107CFU–1013CFU/g, e.g. 108CFU/g–1012CFU/g，109CFU/g-1012CFU/g，2×1010CFU/g-6×1011CFU/g。
According to an embodiment of the invention, the weight ratio of the protective agent to the probiotic may be (0.1-2):1, for example (0.2-1.8):1, (0.3-1.6):1, (0.4-1.5):1, (0.5-1.5): 1.
According to an embodiment of the present invention, the probiotic may be a liquid formulation or a solid formulation. For example, it may be an oral liquid, or a powder, granule or tablet.
On the other hand, the invention also provides a preparation method of the microecological preparation, which comprises the following steps:
step (1): preparing a ginseng extract: extracting Ginseng radix with water under reflux, and concentrating the extractive solution to obtain Ginseng radix extract;
step (2): culturing probiotics: inoculating and culturing the activated probiotics, and then collecting thalli to obtain bacterial sludge;
and (3): and (3) mixing the ginseng extract obtained in the step (1), the bacterial sludge obtained in the step (2) and a protective agent to obtain the liquid microecological preparation.
According to an embodiment of the present invention, the method for preparing a microecological preparation further comprises the step (4) of freeze-drying the liquid microecological preparation to obtain a powdered microecological preparation.
According to an embodiment of the present invention, the ginseng powder is used for the water reflux extraction in the step (1). Adding water 6-12 times, such as 8-10 times of the weight of Ginseng radix, and extracting with water under reflux for 1-4 times, such as 2-3 times.
According to an embodiment of the present invention, the density of the ginseng extract in step (1) at 20 ℃ may be 1.0-1.4g/L, such as 1.01-1.35g/L, 1.02-1.3 g/L.
According to an embodiment of the present invention, wherein the total content of the ginsenosides Rg1 and Re in the ginseng extract is 0.5% or more, the content of the ginsenosides Rb1 is 0.2% or more; preferably, the total content of the ginsenoside Rg1 and Re in the ginseng extract is more than 0.6%, and the content of the ginsenoside Rb1 is more than 0.25%; illustratively, the total content of ginsenoside Rg1 and Re in the ginseng extract is more than 0.75%, and the content of ginsenoside Rb1 is more than 0.31%.
According to the embodiment of the invention, in the step (2), the probiotic bacteria are cultured until the bacterial liquid OD600 is 1-1.5, the culture is stopped, then the bacteria are centrifugally collected at 4 ℃, and the bacteria are washed for 2-3 times by using purified water to obtain bacterial sludge (inverted non-flowing sludge).
According to an embodiment of the invention, the weight ratio of the ginseng extract to the bacterial sludge in step (3) is (0.1-5):1, such as (0.2-2.5):1, (0.3-2):1, (0.4-1.8):1, (0.5-1.5):1, (0.6-1.2):1, etc.
In still another aspect, the present invention also provides the use of the above-mentioned probiotic for the preparation of a food, health food or pharmaceutical product for the prevention and/or treatment of obesity or metabolic diseases.
According to an embodiment of the invention, the metabolic disease is diabetes, in particular type 2 diabetes, hyperlipidemia or fatty liver.
Advantageous effects
According to the microecological preparation containing the ginseng extract and the probiotics, the traditional Chinese medicine components can provide substrates required by growth and synthesis of active effective molecules for the probiotics, the survival rate of the probiotics is improved, in turn, the complex enzyme system in the probiotics further improves the utilization of the traditional Chinese medicine components of the ginseng, the traditional Chinese medicine components and the probiotics can be combined to play a role, compared with the single use of the traditional Chinese medicine preparation or the probiotics, the microecological preparation has the advantages that the regulation effect on intestinal flora is enhanced, the composition of the intestinal flora can be improved, the blood sugar and blood fat metabolism of an organism can be regulated, and the fat accumulation of the organism can be further reduced.
Drawings
Figure 1 shows the effect of the probiotics on the high fat diet induced body weight and visceral fat in obese mice (wherein HFD stands for high fat diet mice; GBS stands for mice fed with the ginseng extract and probiotic mixed probiotics;. indicates P <0.05,. indicates P <0.01,. indicates P <0.001, n ═ 8). A: body weight; b: epididymal fat; c: perirenal fat.
Figure 2 shows the effect of probiotics on the high fat diet induced blood glucose in obese mice. Wherein, A: fasting blood glucose values; b: fasting insulin value; c: insulin resistance index.
Figure 3 shows the effect of probiotics on the high fat diet induced blood lipids in obese mice. Wherein, A: total cholesterol; b: a triglyceride; c: high density lipoprotein; d: low density lipoprotein.
Detailed Description
The microecological preparation of the present invention, and the preparation method and use thereof will be described in further detail with reference to specific examples. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.
Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.
Example 1 preparation of Ginseng extract
Decocting Ginseng radix powder with 6 times of water for 60 min by water extraction, filtering to obtain decoction, and concentrating with rotary evaporator to obtain Ginseng radix extract with density of 1.02g/L at 20 deg.C. In the obtained Ginseng radix extract, the total content of ginsenoside Rg1 and Re is 0.75%, and the content of ginsenoside Rb1 is 0.31%.
Example 2 preparation of a Microecological preparation of extracts of Bacillus subtilis and Panax Ginseng
Inoculating activated Bacillus subtilis, culturing until OD600 of bacteria solution is 1, stopping culturing, centrifuging at 4 deg.C, collecting thallus, washing with purified water for 3 times to obtain bacterial mud (inverted mud), and weighing.
Gelatin, trehalose, mannitol and vitamin C are prepared into aqueous solution of 0.1g/ml of gelatin, 0.2g/ml of trehalose, 0.1g/ml of mannitol and 0.01g/ml of vitamin C, and the aqueous solution is used as protective agent solution.
The bacterial sludge, the protective agent solution and the ginseng extract obtained in the example 1 are mixed according to the weight ratio of 1:0.5:0.5, and after uniform mixing, freeze drying is carried out (wherein the freezing temperature is-50 ℃) so as to finally obtain the powdery microecological preparation. The final viable count of the preparation is about 2 × 1010CFU/g－6×1011CFU/g。
Example 3 preparation of a Microecological preparation of Saccharomyces cerevisiae and Ginseng radix extract
A microecological preparation comprising extracts of Saccharomyces cerevisiae and Panax ginseng was prepared by replacing Bacillus subtilis with Saccharomyces cerevisiae in accordance with the procedure of example 2. Wherein the weight ratio of the ginseng extract to the saccharomyces cerevisiae bacterial mud is 0.6: 1.
Examples of effects
The experimental method comprises the following steps: an obese mouse model is established by adopting high-fat diet, administration treatment is carried out in a gastric lavage mode, indexes such as the weight, the visceral fat weight, the blood sugar and the blood fat of a mouse are detected after continuous intervention for 4 weeks, and specific experimental results are shown in a figure 1, a figure 2 and a figure 3.
FIG. 1 is a graph showing the effects of probiotics on the body weight and visceral fat of high-fat diet-induced obese mice (wherein HFD represents high-fat diet mice; GBS represents mice fed with the ginseng extract and probiotic mixed probiotics prepared in example 2, and the probiotics prepared in example 2 were formulated to have a viable cell dose of 0.2X 108CFU/mL solution, then gavage to mice 0.2 mL; denotes P<0.05, represents P<0.01, represents P<0.001, n ═ 8). A: body weight; b: epididymal fat; c: perirenal fat.
As can be seen from fig. 1, the probiotics can alleviate the weight gain caused by high fat diet and reduce the accumulation of visceral fat.
Figure 2 shows the effect of probiotics on the high fat diet induced blood glucose in obese mice. Wherein, A: fasting blood glucose values; b: fasting insulin value; c: insulin resistance index.
As can be seen from fig. 2, the probiotics significantly reduced fasting blood glucose values, reduced insulin resistance index, improved blood glucose levels and insulin resistance.
Figure 3 shows the effect of probiotics on the high fat diet induced blood lipids in obese mice. Wherein, A: total cholesterol; b: a triglyceride; c: high density lipoprotein; d: low density lipoprotein.
As can be seen in fig. 3, the probiotics significantly reduced triglyceride and high density lipoprotein levels and improved blood lipid levels.
The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. A probiotic, comprising a ginseng extract and a probiotic; wherein the Ginseng radix extract is water extract of Ginseng radix; the probiotic is selected from one or two or more of bifidobacteria, lactobacilli, streptococci, bacilli, clostridium butyricum and yeasts;
preferably, the probiotic bacteria are selected from one, a combination of two or more of lactobacillus acidophilus, lactobacillus rhamnosus, lactobacillus paracasei, lactobacillus casei, lactobacillus johnsonii, lactobacillus plantarum, lactobacillus fermentum, lactobacillus lactis, lactobacillus delbrueckii, lactobacillus helveticus, lactobacillus bulgaricus, lactococcus lactis diacetyl subspecies, lactococcus lactis cremoris, streptococcus thermophilus, bifidobacterium lactis, bifidobacterium animalis, bifidobacterium longum, bifidobacterium breve, bifidobacterium infantis, bifidobacterium adolescentis, lactobacillus reuteri, lactobacillus grignard, bifidobacterium bifidum, bacillus coagulans, bacillus subtilis, bacillus licheniformis, clostridium butyricum, and yeasts;
more preferably, the probiotic is selected from one or a combination of two or more of bifidobacterium lactis, bifidobacterium animalis, bifidobacterium longum, bifidobacterium breve, bifidobacterium infantis, bifidobacterium bifidum, bacillus coagulans, bacillus subtilis and bacillus licheniformis;
most preferably, the probiotic is bacillus subtilis.
2. The microecological preparation according to claim 1, wherein the total content of ginsenoside Rg1 and Re in the ginseng extract is 0.5% or more, and the content of ginsenoside Rb1 is 0.2% or more.
3. The probiotic of claim 1 or 2, wherein the probiotic further comprises a protectant;
preferably, the protective agent is selected from one or a combination of two or more of gelatin, trehalose, mannitol, vitamin C, sodium glutamate, casein, skim milk and sorbitol; more preferably, the protective agent is a mixture of gelatin, trehalose, mannitol and vitamin C, and the weight ratio of the gelatin to the trehalose to the mannitol to the vitamin C is (5-50): (5-100): (5-50): (0.1-2).
4. The probiotic of claim 3, wherein the weight ratio of ginseng extract to probiotic is (0.1-5):1, e.g. (0.2-2.5):1, (0.3-2):1, (0.4-1.8):1, (0.5-1.5):1, (0.6-1.2): 1; the weight ratio of the protective agent to the probiotic is (0.1-2) to 1, such as (0.2-1,8) to 1, (0.3-1.6) to 1, (0.4-1.5) to 1, (0.5-1.5) to 1;
preferably, the viable count of the probiotics in the microecological preparation is about 107CFU–1013CFU/g, e.g. 108CFU/g–1012CFU/g，109CFU/g-1012CFU/g，2×1010CFU/g-6×1011CFU/g。
5. The probiotic formulation according to any one of claims 1 to 4, wherein said probiotic formulation is a liquid formulation or a solid formulation, such as an oral liquid, or a powder, granules or tablets.
6. A method of preparing the probiotic of any of claims 1 to 5, comprising the steps of:
step (1): preparing a ginseng extract: extracting Ginseng radix with water under reflux, and concentrating the extractive solution to obtain Ginseng radix extract;
step (2): culturing probiotics: inoculating and culturing the activated probiotics, and then collecting thalli to obtain bacterial sludge;
and (3): mixing the ginseng extract in the step (1), the bacterial sludge in the step (2) and a protective agent to obtain a liquid microecological preparation; and
optionally, step (4), freezing and drying the liquid microecological preparation obtained in step (3) to obtain a powdery microecological preparation.
7. The preparation method according to claim 6, wherein the ginseng powder is used for water reflux extraction in the step (1); preferably, the water amount is 6-12 times of the weight of Ginseng radix, and reflux extracting for 1-4 times.
8. The method according to claim 6, wherein in the step (2), the probiotic bacteria are cultured until the OD600 of the bacterial liquid is 1-1.5, the culture is stopped, then the bacteria are collected by centrifugation, and the bacteria are washed with purified water for 2-3 times to obtain bacterial sludge.
9. Use of the probiotic according to any one of claims 1 to 5 or of the probiotic obtained according to the process of any one of claims 6 to 8, for the preparation of a food, health food or pharmaceutical product for the prevention and/or treatment of obesity or metabolic diseases.
10. Use according to claim 9, wherein the metabolic disease is diabetes, in particular type 2 diabetes, hyperlipidemia or fatty liver.
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CN112386612A (en) * 2020-12-01 2021-02-23 贾卫国 Probiotic biological preparation for preventing diabetes and preparation process thereof
CN113116941A (en) * 2021-03-08 2021-07-16 山东晶辉生物技术有限公司 Probiotics and prebiotics compound preparation capable of relieving type 2 diabetes and preparation method thereof
CN113897304A (en) * 2021-09-07 2022-01-07 康美药业股份有限公司 Compound strain composition capable of improving ginsenoside conversion degree and application thereof
CN113897304B (en) * 2021-09-07 2024-03-29 康美药业股份有限公司 Composite bacterial strain composition capable of improving ginsenoside conversion degree and application thereof
CN114191463A (en) * 2021-12-07 2022-03-18 鞠懿 New composition of traditional Chinese medicine and probiotics for tonifying qi and preparation method and application thereof
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CN114191363A (en) * 2021-12-30 2022-03-18 哈尔滨求真生物科技有限公司 Antibacterial fermentation product, and preparation method, product and application thereof
CN114703105A (en) * 2022-04-29 2022-07-05 善恩康生物科技（苏州）有限公司 Application of composite probiotics in reducing blood fat or relieving obesity
CN114703105B (en) * 2022-04-29 2022-12-27 善恩康生物科技（苏州）有限公司 Application of composite probiotics in reducing blood fat or relieving obesity
CN115141765A (en) * 2022-05-26 2022-10-04 广州朴成生物科技有限公司 Composite microecological preparation for repairing intestinal tract injury caused by piglet weaning stress and preparation method thereof
CN115141766A (en) * 2022-05-26 2022-10-04 广州朴成生物科技有限公司 Application of composite microecological preparation in preparation of preparation for repairing intestinal tract injury caused by weaning stress of piglets

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